26614 PageRuler Unstained Protein Ladder

Introduction
PRODUCT INFORMATION
Expiry Date: __
WARNING! HARMFUL IF INHALED OR ABSORBED
THROUGH SKIN. CAUSES RESPIRATORY TRACT, EYE
AND SKIN IRRITATION. MAY CAUSE ALLERGIC SKIN
REACTION. MAY BE HARMFUL IF SWALLOWED.
CONTAINS MATERIAL WHICH CAUSES DAMAGE TO THE
FOLLOWING ORGANS: KIDNEYS, LUNGS, UPPER
RESPIRATORY TRACT, SKIN, EYES. CONTAINS MATERIAL
WHICH MAY CAUSE DAMAGE TO THE FOLLOWING
ORGANS: LIVER, GASTROINTESTINAL TRACT.
Avoid exposure - obtain special instructions before use. Do
not breathe vapor or mist. Do not ingest. Do not get in eyes
or on skin or clothing. Use only with adequate ventilation.
Keep container tightly closed and sealed until ready for
use. Wash thoroughly after handling. Refer to MSDS.
Store at -20°C
www.thermoscientific.com/onebio
www.thermoscientific.com/pierce
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Lot: XXXXXXXX
|00072559?~
2 x 250 µl
0026614
#26614
|0026610®~
PageRuler Unstained Protein Ladder
The Thermo Scientific™ PageRuler™ Unstained Protein Ladder
consists of a pre-stained mixture of 14 recombinant, highly
purified proteins ranging from 10 kDa to 200 kDa The ladder is
visualized by SDS-PAGE using coomassie or silver stains or
detected in Western blots with protein stains. For easy reference,
the 50 kDa protein band has a greater intensity than the other
proteins in the ladder. The protein ladder is conveniently
packaged and ready to use with no heating, diluting or additional
reducing agent necessary.
Storage Buffer: 62.5mM Tris•H3PO4 (pH 7.5 at 25°C),
1mM EDTA, 2% (w/v) SDS, 100mM DTT, 1mM NaN3,
0.01% bromophenol blue and 33% (v/v) glycerol.
Important Product Information
• Do not boil the protein ladder.
• In high-percentage gels (14-18%), large proteins
(150-200 kDa) may not separate.
• In low-percentage gels (4-8%), low-molecular weight proteins
may migrate with the dye front.
• The large proteins (> 100 kDa) in the ladder may require longer
transfer times or higher transfer voltages for Western blotting.
• If additional bands appear in the protein ladder, add newly
prepared dithiothreitol (DTT) solution to 100mM final
concentration. DTT oxidation in the storage buffer can cause
the appearance of additional bands.
Rev.5
Procedure for Using the Protein Ladder in
Polyacrylamide Gel Electrophoresis
PageRuler Unstained Protein Ladder
1. Thaw the ladder at room temperature. Do not boil protein
ladder.
2. Mix the solution gently and thoroughly to ensure it is
homogeneous.
3. Load an appropriate volume of the ladder on the gel.
• Mini-gel: 5µL per well (0.75-1.0mm thick) or 10µL per well
(1.5mm thick)
• Large gel: 10µL per well (0.75-1.0mm thick) or 20µL per
well (1.5mm thick)
Note: Dilute the ladder approximately 1/10 in reducing sample
buffer for silver staining.
4. Return the unused protein ladder to -20°C.
Note
To avoid overloading gels which will be subsequently silver
stained, dilute the ladder in protein loading buffer just prior to
use:
Water, nuclease-free (#R0581)
4X Protein Loading Buffer
(e.g. LDS Sample Buffer, Non-Reducing
#R84788, www.thermoscientific.com/pierce)
Reducing Agent
Protein ladder
To final 50 µL volume
12.5 µL
to 50mM final conc.
1 µL
Load 5 µL of the diluted ladder per well for a mini gel/blot and
10 µL per well for a large gel/blot.
8-16% Tris-glycine SDS-PAGE
General References
Burnette, W.N. (1981). “Western blotting”: electrophoretic transfer of
proteins from sodium dodecyl sulfate – polyacrylamide gels to
unmodified nitrocellulose and radiographic detection with antibody and
radioiodinated protein A. Anal Biochem 112(2):195-203.
Kurien, B.T. and Scofield, R.H. (2003). Protein blotting: a review. J Imm
Meth 274:1-15.
Laemmli, U.K. (1970). Cleavage of structural proteins during the
assembly of the head of bacteriophage T4. Nature 227:680-5.
Towbin, H., et al. (1979). Electrophoretic transfer of proteins from
polyacrylamide gels to nitrocellulose sheets: procedure and some
applications. Proc Natl Acad Sci USA 76:4350-4.
This product is manufactured under the license for Strep-tag® technology covered by
US patents Nos. 5,506,121, 6,103,493 and foreign counterparts.
PRODUCT USE LIMITATION
This product is developed, designed and sold exclusively for research purposes and
in vitro use only. The product was not tested for use in diagnostics or for drug
development, nor is it suitable for administration to humans or animals.
Please refer to www.thermoscientific.com/onebio for Material Safety Data Sheet of
the product.
© 2013 Thermo Fisher Scientific, Inc. All rights reserved. All trademarks are the
property of Thermo Fisher Scientific, Inc. and its subsidiaries.

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